Format

Send to

Choose Destination
J Am Chem Soc. 2018 Nov 21;140(46):15701-15711. doi: 10.1021/jacs.8b08050. Epub 2018 Nov 6.

Probing Antibody Binding to Canine Parvovirus with Charge Detection Mass Spectrometry.

Author information

1
Department of Chemistry , Indiana University , 800 E. Kirkwood Ave. , Bloomington , Indiana 47405 , United States.
2
Baker Institute for Animal Health, Department of Microbiology and Immunology, College of Veterinary Medicine , Cornell University , Ithaca , New York 14850 , United States.

Abstract

There are many techniques for monitoring and measuring the interactions between proteins and ligands. Most of these techniques are ensemble methods that can provide association constants and in some cases stoichiometry. Here we use charge detection mass spectrometry (CDMS), a single particle technique, to probe the interactions of antigen binding fragments (Fabs) from a series of antibodies with the canine parvovirus (CPV) capsid. In addition to providing the average number of bound Fabs as a function of Fab concentration (i.e., the binding curve), CDMS measurements provide information about the distribution of bound Fabs. We show that the distribution of bound ligands is much better at distinguishing between different binding models than the binding curve. The binding of Fab E to CPV is a textbook example. A maximum of 60 Fabs bind and the results are consistent with a model where all sites have the same binding affinity. However, for Fabs B, F, and 14, the distributions can only be fit by a model where there are distinct virus subpopulations with different binding affinities. This behavior can be distinguished from a situation where all CPV particles are identical, and each particle has the same distribution of sites with different binding affinities. The different responses to viral heterogeneity can be traced to the Fab binding sites. A comparison of Fab binding to new and aged CPV capsids reveals that a post-translational modification at the binding site for Fab E (M569) probably reduces the binding affinity.

PMID:
30398860
DOI:
10.1021/jacs.8b08050

Supplemental Content

Full text links

Icon for American Chemical Society
Loading ...
Support Center